Features of Cytokine and VEGFA Gene Expression Modified with SARS-CoV-2 Virus in an In Vitro Experiment (Using the Example of the SARS-CoV-2 Vaccine Antigen).

The influence of SARS-CoV-2 antigen on the cytokine-producing function of immune cells was studied. We observed suppression of the production of proinflammatory cytokines by 11-46% relative to the spontaneous level under the influence of SARS-CoV-2 antigen vaccine simulator, as well as when it was co-administered with cortisol (IL-6 by 1.8 times and IFNγ by 1.57 times) compared with control samples. IL-8 production was reduced by 1.72 times relative to its spontaneous level. IL-8 production was reduced by 1.72 times relative to its spontaneous level. Under conditions of SARS-CoV-2 stimulation with the vaccine antigen in vitro, an increase in the relative scaled expression of the VEGFA gene by 2.16 times relative to the spontaneous level was observed, which can be regarded as a model "cytokine storm" scenario. The obtained experimental data verify the ideas about the pathogenetic mechanisms of the COVID-19 and can contribute to the development of new approaches to the correction of its complications.

Septoria blotch epidemic process on spring wheat varieties.

The Septoria blotch of spring wheat leaves and ears is one of the most economically significant infections in the Siberian region. In the control systems of Septoria blotch the main ecologically safe element is resistant varieties, which are designed to slow down the pathogens reproduction rate and slow down or stop the development of the epiphytotic process. The purpose of the work was to clarify the species composition of Septoria blotch pathogens for West Siberian regions and spring wheat varieties, to study the epiphytotic process of Septoria differentially on the leaves and ears of varieties, and to evaluate the activity of seed transmission of Parastagonospora nodorum. Studies were carried out in 2016-2018 according to generally accepted methods. Septoria leaf and ear blotch of spring wheat is widespread in West Siberia and the Trans-Urals, causing a decrease in yield by up to 50 % or more with the deterioration in grain quality. The causative agents of the disease are P. nodorum, Septoria tritici, and P. avenae f. sp. triticae, and the species ratio varied across the regions and varieties, and within plant organs. In Novosibirsk Region, P. nodorum completely dominated; S. tritici was 13.8 times less common; and P. avenae f. sp. triticae was a singleton. In Tyumen Region, the dominance of P. nodorum was disrupted in some geographic locations by S. tritici and P. avenae f. sp. triticae. In Altai Krai, P. nodorum predominated at all points studied; S. tritici and P. avenae f. sp. triticae were found everywhere, but 5.6 and 8.6 times less often, respectively. The study of spring wheat varieties of different origins has not revealed any samples immune to Septoria blotch. A differentiated manifestation of resistance to Septoria leaf and ear disease has been established. Some varieties show complex resistance, combining reduced susceptibility to Septoria leaf and ear disease. Seed infection with P. nodorum in the regions of Siberia reached 7 thresholds and was largely (52.5 %) determined by the August weather conditions. The study of the collection of spring wheat varieties from three Siberian regions has revealed the following trend. Transmission of P. nodorum with the seeds of varieties was the most active (7.6 %) in Novosibirsk Region and somewhat weaker in Omsk Region (5.7 %). The most favorable phytosanitary situation was in Kurgan Region, where varieties transmitted P. nodorum to a low degree (2.1 %), below the threshold.

Western Blotting-Based Quantitative Measurement of Myosin II Regulatory Light Chain Phosphorylation in Small Amounts of Non-muscle Cells.

Myosin II is the main molecular motor in the actomyosin-dependent motility in cells. Phosphorylation of the myosin regulatory light chain (RLC) at Ser19 is a prerequisite for smooth muscle/non-muscle myosin II activation and serves as a biochemical equivalent of myosin II activity. Simultaneous phosphorylation at Thr18 further promotes the myosin II ATPase activity. A number of methods have been developed to measure myosin RLC phosphorylation at Ser19 or di-phosphorylation at Thr18/Ser19. While these methods are straightforward and robust in myosin-rich muscle tissues, they demonstrate limited applicability in non-muscle cells that have low myosin II content and are usually available in lesser amounts than muscle tissue. Because of this, dynamic analysis of RLC phosphorylation in multiple samples of non-muscle cells is difficult and requires large number of cells. The use of phospho-specific antibodies increases detection sensitivity but allows estimation of only relative levels of RLC phosphorylation at specific residues, which makes it difficult to estimate the physiologic relevancy of the observed changes in RLC phosphorylation. To measure RLC phosphorylation in small amounts of non-muscle cells, we used external calibration standards of non-phosphorylated and in vitro phosphorylated RLC in standard SDS-PAGE and Western blot procedures with phospho-specific RLC antibodies. Here, we describe the method in detail and demonstrate its application for quantitative measurement of myosin RLC phosphorylation in endothelial cells in response to natural agonists (thrombin or insulin) and intact human platelets. We discuss the advantages and limitations of the proposed method vs other approaches for measuring myosin RLC phosphorylation in non-muscle cells.

Analysis of the Role of Carriership of Polymorphic Genotypes of ESR1, eNOS, and APOE4 Genes in the Development of Arterial Hypertension in Men.

We studied the role of the carrier status for polymorphic loci of genes encoding estrogen receptors (ESR1), endothelial NO synthase (eNOS), and apolipoprotein E (APOE4) and products of their expression nitrogen oxide (NO) and apolipoprotein (ApoE) in the development of arterial hypertension in men. Conventionally healthy volunteers and 149 men with clinical manifestations of stage I-II arterial hypertension were examined. In men with arterial hypertension, the frequency of minor allele A of ESR1 gene was higher (27.5 vs. 9.5% in the reference group; χ2=4.43, p=0.04). The level of NO in the peripheral blood was also higher in the main group (χ2=3.93, p=0.047). The increase in NO concentration did not depend on the presence of polymorphic genotypes (GG and GT) of eNOS gene, but the decrease in ApoE level in blood serum was associated with TC genotype of APOE4 gene (p=0.04). Our results suggest that minor allele A of ESR1 gene is associated with the development of arterial hypertension in men. Reduced content of ApoE in blood serum of men with arterial hypertension was associated with APOE4 gene polymorphism. However, increased level of NO did not depend on polymorphic genotypes GG and GT of eNOS gene. These polymorphisms are of specific interest as additional markers of genetic predisposition to the development of arterial hypertension in middle-age men.

[Suppression of vascular endothelium hyperpermeability by cell-permeating peptide inhibitors of myosin light chain kinase].

Novel peptides originating from the peptide inhibitor of myosin light chain kinase, L-PIK (Arg-Lys-Lys-Tyr-Lys-Tyr-Arg-Arg-Lys), have been studied for ability to attenuate the thrombin-induced hyperpermeability of endothelial cell monolayer in culture. Peptides [NalphaMeArg1]-Lys-Lys-Tyr-Lys-Tyr-Arg-(D)Arg8-Lys and H-Arg(NO2)Lys-Lys-Tyr-Lys-Tyr-Arg-Arg-Lys-NH2 (designated PIK2 and PIK4, respectively) appeared to be the most effective inhibitors of endothelial cell monolayer hyperpermebility, and surpassed other known peptide inhibitors of myosin light chain kinase derived from original L-PIK. Our results validate PIK2 and PIK4 as the leading molecules for the development of novel drugs intended to counteract pathological hyperpermeability of vascular endothelium.

Interactions between bioactive ferulic acid and fumed silica by UV-vis spectroscopy, FT-IR, TPD MS investigation and quantum chemical methods.

The interactions of bioactive ferulic acid with fumed silica were studied by UV/vis spectroscopy, FT-IR, TPD MS techniques and quantum chemical methods. It was found that surface complexes may form through phenol or carboxyl group of ferulic acid depending on its coverage value. The structure of surface complexes and mechanisms of the ferulic acid chemosorption on the silica surface are discussed. The kinetic parameters of the chemical reactions on silica surface are calculated. The mechanisms of thermal transformations of the ferulic chemosorbed surface complexes are proposed.

Influence of morphology and composition of fumed oxides on changes in their structural and adsorptive characteristics on hydrothermal treatment in steam phase at different temperatures.

A series of fumed oxides such as silica, titania, alumina, silica/alumina (SA), silica/titania (ST), and alumina/silica/titania (AST), initial and hydrothermally treated (HTT) in the steam phase at T(HTT)=150, 250, and 350 degrees C was studied by adsorption, AFM, XRD, FTIR, and theoretical methods. Diminution of the size of primary particles (corresponding to increasing S(BET)) of initial silica and mixed oxides results in enhancement of their structural changes on HTT with elevating T(HTT) and increasing density of packing of primary particles in the secondary structures. Relative changes in the texture of treated fumed silicas are smaller than those of mesoporous silica gels occurring under similar HTT conditions. On HTT, aggregates of primary particles and their agglomerates become denser but their surface layers become looser because of transfer of silica fragments from one particle to another, and the smaller the initial primary particles, the greater the relative diminution of the specific surface area S(BET) for the same type of primary particle packing in aggregates. Relative changes in the pore volume V(p) (or V(BJHd)) on HTT are more complex than that of S(BET), as for many samples the V(p) value increases especially at T(HTT)=150 degrees C. Alumina and titania partially inhibit structural changes on HTT, which decrease in the series silica > SA > AST approximately ST.